The present invention relates to particular marker proteins that can be used in the prognosis of prostate cancer. The present invention further relates to novel transcription factors that can effect apoptosis in cancerous cells. Methods of treating cancer cells, and prostate cancer cells are also provided. The nucleic acid and amino acid sequences of the novel transcription factors are provided along with probes, including nucleotide probes and antibodies, which can be used to determine the presence or absence of the novel transcription factor.
It is generally acknowledged in the medical community that all men will eventually develop prostate cancer, provided that they live long enough for the condition to develop. For example, 50% of all men over 50, and essentially all men over 70 suffer from some form of prostate hyperplasia. Indeed, prostate cancer is the most frequently diagnosed cancer in the United States, with over a quarter of a million new cases being diagnosed each year. Despite the roughly $4 billion dollars per year spent treating this disease, forty thousand men die every year due to prostate cancer, which makes prostate cancer the second leading cause of cancer death in men.
Although the pathogenesis of prostate cancer has not been completely delineated, androgen is believed to play an important role in the development and progression of prostate cancer. It is well established that androgen-dependent growth of the normal prostate stops once the gland reaches the normal size. Indeed, androgen controls the homeostasis of the normal prostate through the androgen action pathway, a cascade of molecular and cellular events triggered by androgen leading to cell growth, differentiation, and/or death. In addition, programmed cell death (i.e., apoptosis) is triggered in the prostate when testosterone levels are completely depleted, and the prostate undergoes regression. Thus, castrated mammals have been employed as an experimental animal model for studying prostate cancer and cDNA collections enriched in genes regulated in prostate homeostasis, and prostrate regression have been disclosed [U.S. Pat. No. 5,821,352, Issued Oct. 13, 1998; and U.S. Pat. No. 5,928,871, Issued Jul. 27, 1999, the contents of which are hereby incorporated by reference in their entireties].
As alluded to above, one characteristic of prostate cancer is that it generally arises relatively late in life and then progresses slowly. If this were always true, the optimal medical response would be to simply monitor the progression of the cancer rather than aggressively treating it, since by the time the cancer progressed to a life threatening stage, the patient would have likely expired due to other more rapidly progressing factors. However, prostate cancers are highly heterogeneous in their progression. Some cancers grow very rapidly and need to be treated aggressively, whereas others are very slow growing and not life-threatening. Thus, one of the most important considerations in the present day treatment of prostate cancer is distinguishing aggressive prostate cancers, which need aggressive treatment, from less aggressive ones, which only require monitoring.
Unfortunately, there are no prognostic tests that can distinguish aggressive prostate cancers from less aggressive forms of the disease. Currently, there is no effective way to distinguish aggressive prostate cancers from slow-growing prostate cancers. Indeed, the present technology relies on monitoring the protein PSA, which not only results in a high percentage of false positives, but also cannot be used as a predictor of the future progression of the disease.
Considering the severe side-effects and expense associated with treating cancer, and prostate cancer treatment in particular, better prognosis tools are desperately needed. Therefore, there is a need to identify other factors that are diagnostic of cancer, and prostate cancer in particular. Furthermore, there is a need to identify means that can be used to accurately predict the progression of cancer, such as prostate cancer. In addition, there is a need to identify means that can be used to identify individual stages of the progression of prostate regression. Furthermore, there is a need to identify factors that can be used in the treatment of cancer, and in particular, prostate cancer.
The citation of any reference herein should not be construed as an admission that such reference is available as xe2x80x9cPrior Artxe2x80x9d to the instant application.
The present invention therefore provides methods that allow aggressive forms of cancer to be identified. In one such embodiment, an aggressive form of prostate cancer is identified. Importantly, a specific protein, TID-1 (otherwise known as TRAITS) is shown to be down-regulated in aggressive forms of cancer. Such cancers include epithelium-derived carcinomas, kidney cancers, lymphomas, leukemias and particularly, prostate cancer.
A particular aspect of the present invention provides the identity of two proteins that are down-regulated in aggressive prostate cancer, but not in slowly progressing prostate cancers. As provided herein, the two prostate proteins, calreticulin and TID-1, are shown to play important roles in the part of the androgen action pathway that suppresses cell proliferation and/or prevents prostate cancer. Therefore the expression of calreticulin (e.g., human calreticulin having the amino acid sequence of SEQ ID NO:36, encoded by the nucleic acid sequence of SEQ ID NO:35) and TID-1 (e.g., human TID-1 having the amino acid sequence of SEQ ID NO:18, encoded by the nucleic acid sequence of SEQ ID NO:17) in prostate cancer cells can be used as markers to distinguish aggressive forms of prostate cancer, which require immediate treatment, from slow growing forms that need only to be monitored.
Although the present invention is not dependent on any particular model, as disclosed below, the present invention is consistent with the unexpected finding that part of the androgen action pathway acts to suppress cell proliferation. This growth suppression is essential for prostate homeostasis and furthermore, limits the cell number in a healthy prostate. In contrast, the inactivation of the part of the androgen action pathway that acts to suppress cell proliferation results in uncontrolled growth leading to prostate cancer.
Therefore, the present invention provides methods of identifying an animal subject, preferably a human subject that is likely to have an aggressive form of prostate cancer. In one embodiment the likelihood determined is between 50 to 100%. In another embodiment the likelihood determined is greater than 70%. In a preferred embodiment, the method identifies an individual that has an 80% or more likelihood of having an aggressive form of prostate cancer. One such method comprises determining the level of calreticulin in a prostate sample from the animal subject. In one embodiment, the sample is obtained by radical prostatectomy. In another embodiment, the sample is obtained by needle biopsy.
When the level of calreticulin determined is 75% or more down-regulated in tumor cells relative to that determined in benign prostatic epithelial cells of the same specimen, the animal subject is identified as being likely to have an aggressive form of prostate cancer. In one embodiment the determination of the level of calreticulin is performed in situ. In another embodiment the determination of the level of calreticulin is performed in vitro. In still another embodiment, the determination of the level of calreticulin is performed in vivo. In a preferred embodiment, the determination of the level of calreticulin is performed by Laser Capture Microscopy coupled with a Western blot.
In a particular embodiment the determination of the level of calreticulin is performed with an antibody specific for calreticulin. In another such embodiment the determination of the level of calreticulin is performed by PCR with a primer specific for an mRNA encoding calreticulin. In still another embodiment the determination of the level of calreticulin is performed with a nucleotide probe specific for an mRNA encoding calreticulin. In one such embodiment, the determination of the level of calreticulin is performed by a Northern blot. In another embodiment, the determination of the level of calreticulin is performed by a ribonuclease protection assay.
In a related embodiment, the method further comprises detecting the TID-1 in a prostate sample from the animal subject. The TID-1 can be detected by Western blot and/or Northern blot and/or as provided for below. When the TID-1 is low to undetectable in the prostate sample, (e.g., undetectable being not detectable by a Western blot and/or Northern blot and/or immunohistochemistry) the subject is further confirmed as being likely to have an aggressive form of prostate cancer.
In yet another embodiment, the method comprises detecting the TID-1 in a prostate sample from the animal subject without determining the level of calreticulin. When the TID-1 is low to undetectable in a tumor cell of a prostate sample (e.g., no staining by immunohistochemistry is observed or no detection by a Northern blot), the subject is identified as being likely to have an aggressive prostate cancer.
In still another embodiment, the method comprises detecting the TID-1 in a tissue sample from the animal subject. Such tissues include epithelium tissue, kidney, lymph nodes, and blood tissue. When the TID-1 is low to undetectable in a tumor cell of a tissue sample (e.g., no staining by immunohistochemistry is observed or no detection by a Northern blot), the subject is identified as being likely to have an aggressive cancer.
The present invention also provides methods of identifying an animal subject that is likely to have a slow growing prostate cancer. In one embodiment the likelihood determined is between 50 to 100%. In another embodiment the likelihood determined is greater than 70%. In a preferred embodiment, the method identifies an individual that has an 80% or more likelihood of having a slow growing form of prostate cancer. One such embodiment comprises detecting TID-1 in a prostate sample from the subject. The TID-1 can be detected by Western blot and/or Northern blot and/or as provided for below. When the TID-1 is detectable in the prostate sample from the animal subject the animal subject is identified as being likely to have a slow growing prostate cancer. In one embodiment, the sample is obtained by radical prostatectomy. In another embodiment, the sample is obtained by needle biopsy. In a particular embodiment the level of TID-1 detected is about 50% or more compared to the normal level observed in slow growing prostate cancers.
In one embodiment the determination of the level of TID-1 is performed in situ. In another embodiment the determination of the level of TID-1 is performed in vitro. In still another embodiment, the determination of the level of TID-1 is performed in vivo. In a particular embodiment the determination of the level of TID-1 is performed with an antibody specific for TID-1. In another such embodiment the determination of the level of TID-1 is performed by PCR with a primer specific for an mRNA encoding TID-1. In still another embodiment the determination of the level of TID-1 is performed with a nucleotide probe specific for an mRNA encoding TID-1. In still another embodiment the determination of the level of TID-1 is performed by a Northern blot. In yet another embodiment the determination of the level of TID-1 is performed by a ribonuclease protection assay. In still another embodiment the determination of the level of TID-1 is performed by immunohistochemistry. In yet another embodiment the determination of the level of TID-1 is performed by Laser capture microscopy coupled with a Western blot. In still another embodiment the determination of the level of TID-1 is performed by RT-PCR.
In a preferred embodiment, the method further comprises determining the level of calreticulin in a prostate sample from the subject. When the level of calreticulin is not down-regulated e.g., no more than about 50% down-regulated in the tumor cells relative to that determined in benign prostatic epithelial cells of the same specimen, the subject is identified as being likely to have a slow growing prostate cancer.
In yet another embodiment, the level of calreticulin in a prostate sample from the subject is determined without determining the TID-1. When the level of calreticulin is not down-regulated in the prostate sample from the subject relative to a healthy prostate sample, the subject is identified as being likely to have a slow growing prostate cancer.
In another aspect of the present invention an isolated nucleic acid encoding a TID-1 is provided. In one such embodiment, the nucleic acid encodes a TID-1 that is a transcription factor comprising an amino acid sequence that has at least 25% identity with that of SEQ ID NO:18. In a preferred embodiment, the nucleic acid encodes a TID-1 that comprises a nuclei localization signal and/or a glutamine rich region. Preferably, the nucleic acid encodes a TID-1 that is localized in the nuclei. In a particular embodiment the nucleic acid encodes a TID-1 whose expression is restricted to the male sex accessory organs, as is the case in the rat. In another embodiment, the nucleic acid encodes a TID-1 that has an apoptosis-inducing domain (e.g., the protein and/or a fragment thereof can induce apoptosis in a cell). In another embodiment, the nucleic acid encodes a TID-1 that has a transactivation domain. Preferably, the nucleic acid encodes a TID-1 whose expression is regulated by testosterone.
In a preferred embodiment, the nucleic acid encodes a TID-1 that is a mammalian protein. In embodiment of this type, the nucleic acid encodes a rat TID-1 protein. In one such embodiment, the rat protein comprises the amino acid sequence of SEQ ID NO:14. In a particular embodiment of this type, the nucleic acid that encodes the rat TID-1 comprises the nucleotide sequence of SEQ ID NO:13. In another such embodiment the nucleic acid encodes a rat protein that comprises the amino acid sequence of SEQ ID NO:14 comprising a conservative amino acid substitution.
In another embodiment the nucleic acid encodes a mouse TID-1 protein. In one such embodiment, the mouse protein comprises the amino acid sequence of SEQ ID NO:16. In a particular embodiment of this type, the nucleic acid that encodes the mouse TID-1 comprises the nucleotide sequence of SEQ ID NO:15. In another such embodiment the nucleic acid encodes a mouse protein that comprises the amino acid sequence of SEQ ID NO:16 comprising a conservative amino acid substitution.
In yet another embodiment the nucleic acid encodes a human TID-1 protein. In one such embodiment, the human protein comprises the amino acid sequence of SEQ ID NO:18. In a particular embodiment, the nucleic acid that encodes the human TID-1 comprises the nucleotide sequence of SEQ ID NO 17. In another such embodiment the nucleic acid encodes a human protein that comprises the amino acid sequence of SEQ ID NO:18 comprising a conservative amino acid substitution.
In a related embodiment, the nucleic acid encodes a mammalian EAF1 protein. In a preferred embodiment the mammalian EAF1 is a human protein. In one such embodiment, the human protein comprises the amino acid sequence of SEQ ID NO:20. In a particular embodiment of this type, the nucleic acid that encodes the human EAF1 comprises the nucleotide sequence of SEQ ID NO:19. In another such embodiment the nucleic acid encodes a human EAF1 that comprises the amino acid sequence of SEQ ID NO:20 comprising a conservative amino acid substitution.
The present invention further provides nucleic acids encoding a functional fragment of TID-1, e.g., the apoptosis-inducing domain and/or the transactivation domain. In a preferred embodiment the nucleic acid encodes an apoptosis-inducing domain comprising the 46 amino acid residues encoded essentially by exon III of a TID-1 (see FIGS. 14 and 18). In an alternative embodiment the transactivation domain comprises the 147-149 amino acids encoded by exons IV-VI of a TID-1 (see FIGS. 14 and 18).
In a particular embodiment, the nucleic acid encodes amino acids 1-113 of SEQ ID NO:14. In another embodiment, the nucleic acid encodes amino acids 1-113 of SEQ ID NO:16. In still another embodiment, the nucleic acid encodes amino acids 1-113 of SEQ ID NO:18. In yet another embodiment, the nucleic acid encodes amino acids 68-113 of SEQ ID NO:14. In still another embodiment, the nucleic acid encodes amino acids 68-113 of SEQ ID NO:16. In yet another embodiment, the nucleic acid encodes amino acids 68-113 of SEQ ID NO:18. In a preferred embodiment, the nucleic acid encodes 8 to 40 (more preferably 15 to 25) consecutive amino acids from amino acid residues 68-113 of SEQ ID NO:18. Preferably, the product expressed by the nucleic acid retains the ability to stimulate apoptosis in a cell.
All of the nucleic acids of the present invention can further comprise a heterologous nucleotide sequence. In addition, recombinant DNA molecules that are operatively linked to an expression control sequence can be constructed from and/or derived from the nucleic acids of the present invention. Furthermore, expression vectors containing the recombinant DNA molecules of the present invention are also provided. In addition, cells that have been transfected and/or transformed with the expression vectors of the present invention, in which the TID-1 or EAF1 protein is expressed by the cell are also part of the present invention. In a preferred embodiment, the cell is a mammalian cell.
The present invention also provides methods of expressing the recombinant TID-1 polypeptides and fragments thereof of the present invention in cells containing the expression vectors of present invention. One such method comprises culturing the cell in an appropriate cell culture medium under conditions that provide for expression of the recombinant polypeptide (e.g., TID-1 or EAF1) by the cell. In a preferred embodiment, the method further comprises the step of purifying the recombinant TID-1 or EAF1. The purified form of the recombinant TID-1 or EAF1 is also part of the present invention.
The present invention further provides nucleic acids that hybridize under standard conditions to a nucleic acid of the present invention. In a particular embodiment, the nucleic acid comprises the nucleotide sequence of SEQ ID NO:13. In another embodiment, the nucleic acid comprises the nucleotide sequence of SEQ ID NO:15. In yet another embodiment, the nucleic acid comprises the nucleotide sequence of SEQ ID NO:17. In a preferred embodiment, the nucleic acid encodes a TID-1 that comprises a nuclei localization signal and/or a glutamine rich region. Preferably, the nucleic acid encodes a TID-1 that is localized in the nuclei. In a particular embodiment the nucleic acid encodes a TID-1 whose expression is restricted to the male sex accessory organs, as is the case in the rat. In another embodiment, the nucleic acid encodes a TID-1 that has an apoptosis-inducing domain (e.g., the protein and/or a fragment thereof can induce apoptosis in a cell). In another embodiment, the nucleic acid encodes a TID-1 that has a transactivation domain. Preferably, the nucleic acid encodes a TID-1 whose expression is regulated by testosterone.
The present invention also provides nucleotide probes for all of the nucleic acids of the present invention.
The present invention further provides isolated TID-1 polypeptides and fragments thereof. In one such embodiment, the TID-1 is a transcription factor having an amino acid sequence that has at least 25% identity with that of SEQ ID NO:18. In a preferred embodiment, the TID-1 comprises a nuclei localization signal and/or a glutamine rich region. Preferably, the TID-1 is localized in the nuclei. In a particular embodiment the TID-1 is restricted to the male sex accessory organs, as is the case in the rat. In another embodiment, the TID-1 that has an apoptosis-inducing domain (e.g., the protein and/or a fragment thereof can induce apoptosis in a cell). In another embodiment, the TID-1 that has a transactivation domain. Preferably, the TID-1 whose expression is regulated by testosterone.
In a preferred embodiment, the isolated TID-1 is a mammalian protein. In one such embodiment, the isolated TID-1 is a rat TID-1 protein. In a particular embodiment of this type, the rat protein comprises the amino acid sequence of SEQ ID NO:14. In another such embodiment the rat TID-1 comprises the amino acid sequence of SEQ ID NO:14 comprising a conservative amino acid substitution. In another particular embodiment the isolated TID-1 is a mouse TID-1 protein. In one such embodiment of this type, the mouse TID-1 comprises the amino acid sequence of SEQ ID NO:16. In another such embodiment the mouse TID-1 comprises the amino acid sequence of SEQ ID NO:16 comprising a conservative amino acid substitution. In yet another particular embodiment the isolated TID-1 is a human TID-1 protein. In one such embodiment of this type, the human TID-1 comprises the amino acid sequence of SEQ ID NO:18. In another such embodiment the human TID-1 comprises the amino acid sequence of SEQ ID NO:18 comprising a conservative amino acid substitution.
In a related embodiment, the present invention provides a human EAF1 protein. In one such embodiment, the human protein comprises the amino acid sequence of SEQ ID NO:20. In a particular embodiment of this type, the human EAF1 comprises the amino acid sequence of SEQ ID NO:20 comprising a conservative amino acid substitution.
The present invention further provides functional fragments of the proteins of the present invention, e.g., the apoptosis-inducing domain and the transactivation domain of TID-1. In a preferred embodiment the apoptosis-inducing domain comprises the 46 amino acid residues encoded essentially by exon III of a TID-1 (see FIGS. 14 and 18). In an alternative embodiment the transactivation domain comprises the 147-149 amino acids encoded by exons IV-VI of a TID-1 (see FIGS. 14 and 18).
In a particular embodiment of this type, the fragment comprises amino acids 1-113 of SEQ ID NO:14. In another embodiment, the fragment comprises amino acids 1-113 of SEQ ID NO:16. In still another embodiment, the fragment comprises amino acids 1-113 of SEQ ID NO:18. In yet another embodiment, the fragment comprises amino acids 68-113 of SEQ ID NO:14. In still another embodiment, the fragment comprises amino acids 68-113 of SEQ ID NO:16. In yet another embodiment, the fragment comprises amino acids 68-113 of SEQ ID NO:18. In a preferred embodiment, the fragment comprises 8 to 40 (more preferably 15 to 25) consecutive amino acids from amino acid residues 68-113 of SEQ ID NOs:14, 16, and/or 18. Preferably, the fragment retains the ability to stimulate apoptosis in a cell.
In a particular embodiment, the transactivation domain comprises amino acid residues 114-262 of SEQ ID NOs:14 and 16. In still another embodiment the transactivation domain comprises amino acid residues 114-260 of SEQ ID NOs:18. All of the fragments of the present invention can also contain a conservative amino acid substitution.
The present invention also provides antigenic fragments of the TID-1 and EAF1 polypeptides of the present invention, as well as proteolytic fragments of the TID-1 and EAF1 polypeptides of the present invention. Preferably the proteolytic fragments are eight amino acids or larger. In addition, the present invention further provides chimeric/fusion proteins/peptides comprising the TID-1 polypeptides, and fragments thereof, including functional, proteolytic and antigenic fragments. Moreover, the present invention provides chimeric/fusion proteins/peptides comprising the EAF1 polypeptides, and fragments thereof, including functional, proteolytic and antigenic fragments.
Antibodies to the TID-1 polypeptides, to the chimeric/fusion proteins comprising the TID-1 polypeptides, as well as to the fragments of the TID-1 polypeptides, including proteolytic, and antigenic fragments, and to the chimeric/fusion proteins/peptides comprising these fragments are also part of the present invention as are the corresponding antibodies raised against EAF1 polypeptides and fragments etc. In addition, methods of using such antibodies for the prognosis of cancer, and prostate cancer in particular, are also part of the present invention.
In addition, antibodies to calreticulin are also provided. As above, methods of using such antibodies for the prognosis of prostate cancer are also part of the present invention.
The antibodies of the present invention can be polyclonal antibodies, monoclonal antibodies and/or chimeric antibodies. Immortal cell lines that produce a monoclonal antibody of the present invention are also part of the present invention.
The present invention further provides a non-human knockout animal comprising a disruption in an endogenous allele encoding TID-1. Preferably the disruption prevents the expression of a functional TID-1 from that allele. In a preferred embodiment, the non-human knockout animal further comprises a disruption in a second endogenous allele encoding TID-1. The disruption of both alleles preferably prevents the non-human knockout animal from expressing functional endogenous TID-1. In a preferred embodiment the knockout animal is a mouse having the propensity for having cancer.
The present invention also provides a non-human knockout animal comprising a disruption in an endogenous allele encoding EAF1. Preferably the disruption prevents the expression of a functional EAF1 from that allele. In a preferred embodiment, the non-human knockout animal further comprises a disruption in a second endogenous allele encoding EAF1. The disruption of both alleles preferably prevents the non-human knockout animal from expressing functional endogenous EAF1. In a preferred embodiment the knockout animal is a mouse having the propensity for having cancer.
The present invention also provides a non-human transgenic animal that has been constructed to express additional copies of the TID-1 and/or the EAF1 protein. In a preferred embodiment of this type, the non-human transgenic animal is a mouse.
The present invention further provides methods of inducing cells to undergo apoptosis. One such method comprises administering a TID-1 to the cell. Another such method comprises administering the N-terminal fragment of TID-1 or a portion thereof to the cell. In a particular embodiment of this type the N-terminal fragment comprises amino acids 1-113 of SEQ ID NO:14. In another embodiment, the N-terminal fragment comprises amino acids 1-113 of SEQ ID NO:16. In still another embodiment, the N-terminal fragment comprises amino acids 1-113 of SEQ ID NO:18. In yet another embodiment, the portion of the N-terminal fragment comprises amino acids 68-113 of SEQ ID NO:14. In still another embodiment, the portion of the N-terminal fragment comprises amino acids 68-113 of SEQ ID NO:16. In yet another embodiment, the portion of the N-terminal fragment comprises amino acids 68-113 of SEQ ID NO:18. In a preferred embodiment, the portion of the N-terminal fragment comprises 8 to 40 (more preferably 15 to 25) consecutive amino acids from amino acid residues 68-113 of SEQ ID NOs:14, 16, and/or 18.
In another embodiment the method comprises administering EAF1 to the cell. In a related embodiment of this type a portion of an N-terminal fragment of EAF1 is administered to the cell.
The present invention also includes a method of treating cancer in an animal subject, such as an epithelium-derived carcinoma, a kidney cancer, a lymphoma, a leukemia, and preferably prostate cancer. One such method comprises inducing the expression of TID-1 activity in the cells of a specific tissue (e.g., prostate cells) in a patient. In a particular embodiment the expression of TID-1 activity is induced by providing exogenous TID-1 or a fragment thereof to the subject via gene therapy. In another embodiment the expression of TID-1 activity is induced by peptide-directed delivery of TID-1 protein.
Thus, the present invention provides methods of administering TID-1, an N-terminal fragment of TID-1 or a portion thereof, in vivo to a cancerous cell in an animal subject. In a particular embodiment of this type the N-terminal fragment comprises amino acids 1-113 of SEQ ID NO:14. In another embodiment, the N-terminal fragment comprises amino acids 1-113 of SEQ ID NO:16. In still another embodiment, the N-terminal fragment comprises amino acids 1-113 of SEQ ID NO:18. In yet another embodiment, the portion of the N-terminal fragment comprises amino acids 68-113 of SEQ ID NO:14. In still another embodiment, the portion of the N-terminal fragment comprises amino acids 68-113 of SEQ ID NO:16. In yet another embodiment, the portion of the N-terminal fragment comprises amino acids 68-113 of SEQ ID NO:18. In a preferred embodiment, the portion of the N-terminal fragment comprises 8 to 40 (more preferably 15 to 25) consecutive amino acids from amino acid residues 68-113 of SEQ ID NOs:14, 16, and/or 18. Preferably, the animal subject is a human.
In another embodiment the method of treating comprises administering EAF1 in vivo to a cancerous cell contained by the animal subject. In a related embodiment of this type an N-terminal fragment of EAF1 is administered in vivo to a cancerous cell of the animal subject. Preferably, the animal subject is a human. EAF1 can be administered by the same methods as outlined above for TID-1.
Accordingly, it is a principal object of the present invention to provide new methods in the prognosis of cancer in humans.
It is a further object of the present invention to provide new protein markers that can be used in the prognosis of cancer in humans, e.g., in the prognosis of prostate cancer.
It is a further object of the present invention to provide new transcription factors that are involved in apoptosis.
It is a further object of the present invention to provide specific probes for assaying tumor tissue to determine whether the tumor cells express TID-1.
It is a further object of the present invention to provide methods for assaying prostate tumor tissue to determine whether calreticulin has been down-regulated.
It is a further object of the present invention to provide new treatments for cancer, particularly prostate cancer.
It is a further object of the present invention to provide new agents for stimulating apoptosis in a cell.
It is a further object of the present invention to provide new agents for inhibiting/preventing apoptosis in a cell.
These and other aspects of the present invention will be better appreciated by reference to the following drawings and Detailed Description.